Ultrasensitive CRISPR-based diagnostic for field-applicable detection of <i>Plasmodium</i> species in symptomatic and asymptomatic malaria
Rose Lee, Helena de Puig, Peter Q. Nguyen, Nicolaas M. Angenent-Mari, Nina M. Donghia, James P. McGee, Jeffrey D. Dvorin, Catherine M. Klapperich, Nira R. Pollock, James J. Collins
Abstract
Significance Detection of submicroscopic malaria in asymptomatic individuals is needed for eradication and remains a diagnostic gap in resource-limited settings. Nonfalciparum clinical diagnostics are a second gap, as these infections have a low parasite density and are commonly undetected. We describe an integrated, 60-min, ultrasensitive and specific CRISPR-based diagnostic for the four major pathogenic Plasmodium species that can fill these gaps. Using the SHERLOCK (specific high-sensitivity enzymatic reporter unlocking) platform, we designed assays with limits of detection below that recommended by the World Health Organization. These assays have a simplified sample preparation method: the SHERLOCK parasite rapid extraction protocol, which eliminates complicated nucleic acid extraction steps. Our work further translates the SHERLOCK platform into a field-deployable diagnostic.